Monitoring viral decontamination procedures with green fluorescent protein-expressing adenovirus

Abstract

Recombinant adenoviruses are widely used in life science laboratories and represent an excellent tool for gene transfer to a great variety of cells [1]. Therefore, potential exposure of laboratory personnel to the infectious virions is a serious concern. Here we describe a method combining green Xuorescent protein-expressing adenoviruses (Ad– GFP) with Xuorescence-activated cell sorting (FACS) and Xuorescence microscopy for monitoring virus stability and elimination procedures. Using this method, we have analyzed typical experimental situations and virus elimination protocols commonly applied in life science laboratories. The described method is sensitive and cost-eVective and can be applied to a wide range of experimental conditions. A key advantage of adenoviral gene transfer systems is the possibility to eYciently transduce most human cell lines and primary cells, including nondividing cells such as primary hepatocytes and neurons. An important issue is precluding the exposure of laboratory personnel to the infectious particles [2]. Compared with the situation in manufacturing facilities, cells transduced with recombinant adenoviruses in life science laboratories usually are subjected to a greater variety of downstream procedures. Therefore, it is crucial to know how stable the infectious particles are under diVerent experimental conditions and to what extent frequently used decontamination procedures eliminate infectivity. However, in contrast to data on con-